尖锐湿疣患者外周血自然杀伤细胞活性的研究  

Studies on the Activation of NK Cell from Human Peripheral Blood of the Patients with Condyloma Acuminate

作　　者：李振[1] 曾耀英[2] 蔡小嫦[2] 何贤辉[2] 徐丽慧[2] 曾山[2] 狄静芳[2] 

机构地区：[1]深圳市龙岗中心医院皮肤科,广东深圳518116 [2]暨南大学组织移植与免疫中心,广东广州510632

出　　处：《中国皮肤性病学杂志》2003年第3期145-147,共3页The Chinese Journal of Dermatovenereology

基　　金：国家重点基础研究发展规划项目 ("973")基金资助(G1 9990 5430 3)

摘　　要：目的　从自然杀伤细胞 (NK)内分泌γ 干扰素和CD6 9表达等层次 ,探讨尖锐湿疣 (CA)患者的细胞免疫状态 ,为临床免疫治疗提供指引。方法　用密度梯度离心法分离出外周血单个核细胞 (PBMC) ,进行细胞培养和免疫荧光染色 :①PDB刺激PBMC ,2h、12h、2 4h后收获细胞 ,进行anti CD5 6 FITC与anti CD6 9 PE免疫标记 ;②PDB、Ionomycin和Monensin刺激PBMC ,4h收获细胞 ,用anti CD5 6 PE与anti IFN γ FITC细胞内染色 ,最后利用流式细胞仪对标记细胞进行分析。结果　NK细胞受体外刺激后 ,长病程组各时点的CD6 9阳性细胞百分率分别是 ( 2 5 .73± 2 .99) %、( 5 7.0 8±4.74) %、( 6 7.0 7± 2 .5 2 ) % ,与对照组各时点相比较 ,差异均有显著性 (P <0 .0 5 ) ,而短病程组各时点与对照组比较 ,差异均无显著性 (P >0 .0 5 )。长病程组的IFN γ阳性NK细胞比率是 ( 34.6 9± 3 .36 ) % ,与对照组比较差异有显著性 (P <0 .0 5 )。短病程组与对照组比较差异无显著性 (P >0 .0 5 )。结论　短病程患者NK细胞活化和功能均正常 ,但长病程者NK细胞活化明显不足和功能严重障碍。本试验结果为长病程患者的免疫治疗提供理论依据和新的指引。Objective To improve the clinical therapy,studies were performed on the details of intracellular IFN-γ secretion and CD69 expression on NK cell in vitro for investigating the statute of cellular immunity of the patients with condyloma acuminate(CA).Methods After isolated the PBMC from anti-clotting human peripheral blood with heparin by Ficoll-Hypaque centrifugation,the PBMC culture procedure in vitro and two-color immunofluorescent staining were undertaken as following:①PBMC were activated 2,12,24 hours by PDB,then stained with anti-CD56-FITC and anti-CD69-PE.②PBMC were stimulated 4 hours by PDB,Ionomycinand and Monensin together,then intracellular stained with anti-CD56-PE and anti-IFN-γ-FITC.At last the stained cells were analyzed by flow cytometry.Results The percentages of NK-cells expressing CD69 of long history-CA group at 2,12,24 hour were(25.73±2.99)%,(57.08±4.74)%,(67.07±2.52)% respectively,the differences between them and those of control group were significant(P<0.05)But the differences between the short history-CA group and those of control group were no significant(P>0.05),The ratios of IFN-γ positive NK-cells of long history-CA group were (34.69±3.36)%,the difference between them and control group was significant(P<0.05).But there were no difference between the short history-CA group and those of control group (P>0.05).Conclusion The activation and function of NK cells of the patients with short history-CA were normal,but those of the patients of long history-CA were deficient.These data would provide the theory and new ways for the immune therapy of especially long history-CA.

关 键 词：尖锐湿疣 外周血 自然杀伤细胞活性 γ—干扰素 CD69 细胞免疫状态 临床免疫治疗 

分 类 号：R752.53[医药卫生—皮肤病学与性病学]